Product is the lyophilized powder of goat antiserum to mouse complement C3 and buffer salts.
Background Information
The complement system provides innate defense against microbial infection and is a "complement" to antibody mediated immunity. The complement system consists of thirty-five interacting plasma and membrane associated proteins which contribute to host-defense and initiate and amplify inflammation, even in the preimmune state where specific antibodies and lymphocytes are not available. In addition to the complement components themselves, this system also contains several soluble factors that prevent spontaneous complement activation from occurring in solution, as well as, several regulatory membrane associated proteins that protect host cells from accidental complement attack. Cleavage of the C3 component by either the classical pathway or the alternative pathway releases C3a and C3b. C3a appears to be important in many inflammatory responses while the C3b fragment covalently binds to the cell or bacterial surface and plays a role in opsonisation. Binding of C3b to the C4b component of the C3 convertase, results in C5 convertase (C4b3b2a) formation.
Applications
This product is suitable for use as a precipitating antibody. It gives sharp precipitin lines in immunoelectrophoresis and immunodiffusion techniques. It has been successfully used in immunocytochemistry, western blot, Immunoelectrophoresis, and flow cytometry applications.
Recommended Use
Reconstitute product with 2.0 mL of deionized or distilled water. Gentle swirling may be used to speed rehydration. Avoid vigorous shaking of the reconstituted material.
Preparation Method
The antiserum is prepared by hyperimmunizing goats with highly purified mouse C3; antiserum is delipidated and made specific by solid-phase absorption. The resulting antiserum is dialyzed into 0.02M sodium phosphate, 0.14M sodium chloride, pH 7.3, with 0.05% sodium azide, adjusted to standard titer, filtered through a 0.22 µm filter, vialed and lyophilized.
Analysis Note
The total protein is measured using the Biuret procedure, with bovine albumin as a standard. Antibody titer is standardized by equivalence-point precipitation. Each antiserum is tested for specificity at a minimum of 80 mg/mL using immunoelectrophoresis; Antibody activity to other serum proteins is not present.